Search

The main cause of hepatitis C is hepatitis C virus or HCV and for the cure of hepatitis C, NS3/4A protease has been found an important and emerging target. A number of HCV NS3/4A protease inhibitors have been discovered which have shown subsequent reduction in reducing the viral load leading to this infection however they are still undergoing clinical trials for improvement. Bacterial proteases are of great pharmaceutical importance and have a key role in various biological processes and in life cycle of several pathogens. The current study was planned to explore hexapeptides derived from conserved regions of bacterial proteases for their potential in blocking the NS3 protease activity of HCV which would finally inhibit HCV multiplication. For this, a novel protease gene nprB was isolated from a thermophilic bacterium Streptomyces thermovulgaris and bioinformatics analyses were performed. PCR amplification and sequencing of nprB gene indicated an open reading frame of 178 aa (20191.18 Dalton).The peptide GGVHIN was the top ranked with minimum S-score of -17.21) followed by hexapeptides VDAHAN, GVGREA, GALNES and VHINSS with their S-scores of -14.73, -13.78, -10.72 and -10.70, respectively. A phylogram was also reconstructed to reveal evolutionary relationships of nprB with its various homologs. The provided data will serve as a background to further reveal pharmaceutical and biotechnological importance of this novel protease gene from S. thermovulgaris in future.

Bacterial proteases are of great pharmaceutical importance and have a key role in various biological processes and in life cycle of several pathogens. New technology used for rational protein engineering as well improved delivery options will expand the potential pharmaceutical applications of proteases. The catalytic proteases belong to metalloproteases (EC.3.4.24) that comprise thermo lysine. The metalloproteases and their homologs have many important biotechnological and therapeutic applications. In the present study, a novel protease gene nprB was isolated from a thermophilic bacterium Streptomyces thermovulgaris and bioinformatics analyses were performed. PCR amplification and sequencing of nprB gene indicated an open reading frame of 178 aa (20191.18 Dalton). Based on protein sequence homology as well as conserved motifs and PTF domain the protein is characterized as a thermo lysine-like protease and is a member of M4 family of metalloproteases. Different bioinformatics tools such as ProtParam, SOPMA, signalP4.1 and ProDom from the ExPAsy server were used for structural and functional analyses. A phylogram was also reconstructed to reveal evolutionary relationships of nprB with its various homologs. The provided data will serve as a background to further reveal pharmaceutical and biotechnological importance of this novel protease gene from S. thermovulgaris in future.

Effect of solvent polarity and extraction time on antioxidant properties of Brassica oleracea seeds was optimized by response surface methodology using central composite design. A significant linear negative effect of solvent polarity on total extractable components (TEC), total phenolic acids (TPA), linoleic acid reduction capacity (LARC) and hydroxyl radical scavenging capacity (HRSC) and significant positive effect on iron chelating activity (ICA), reducing power (RP) and (DPPH RSC). Linear effect of solvent polarity on total antioxidant activity (TAOA) was also found to be positive but not significant. Significant linear negative effect of extraction time was observed on ICA and HRSC. Only RP was found to be significantly increased with increase in extraction time. Quadratic effect of solvent polarity was found to be negative on TAOA, DPPH RSC and HRSC. However, quadratic effects of both variables were found to be positive on TEC, TPA, ICA and LARC. The polarity dependent increase in TAOA indicates the extraction of phytochemicals with comparatively stronger antioxidant properties in polar solvents. The decrease in antioxidant properties in response to increase in extraction time may be attributed to the time dependent loss of antioxidant activity of extracted compounds.

The aim of this study was to determine antioxidant activity of Brassica oleracea (Cabbage), Brassica rapa (Turnip) and Raphanus sativus (Radish). All of the three selected vegetables were ultrasonically extracted in water and acetone. The antioxidant activity was determined by DPPH and H2O2. Highest phenolic, flavonoid and flavonol contents were found among water extract of Raphanus sativus, Brassica oleracea and Brassica rapa respectively. Highest %age of DPPH inhibition was exhibited by water extract of Brassica rapa and highest OH ◦ radical scavenging activity was shown by water extract of Raphanus sativus.

Control of harmful bacteria in food, aquaculture, pharmaceuticals, agriculture, hospitals and recreation water pools are of great global concern. Marine bacteria are an enormous source of bio-controlling agents. The aim of this study was to identify and optimize the growth conditions including effect of different biotic and abiotic factors on antimicrobial activity of strain DK1-SA11 isolated from Qingdao Bay of China Yellow Sea. Microscopic characterization, API® 20E and 50 CHB kit base carbohydrates utilization, 16S rDNA and DNA gyrB gene sequencing studies identified the bacterium as Bacillus subtilis subsp. spizizenii DK1-SA11. Antimicrobial spectrum of cell free supernatant (CFS) has shown antimicrobial activities against all test strains including methicillin-resistant Staphylococcus aureus, E. coli O157:H7, Candida albicans, Klebsiella pneumoniae, Listeria monocytogenes, Vibrio parahaemolyticus, E. coli, Pseudomonas fluorescens, Salmonella typhimurium and Vibrio cholerae. Among all the media tested, Marine Broth 2216 was found to be the best medium for bacterial growth and production of antibacterial compounds. The other optimum conditions for growth were pH:7 and incubation temperature: 25°C with < 180 rpm for 60-72 h. Out of 49 different carbohydrates tested, D-mannose increases the antibacterial activity by 33.3% while D-arabitol decreases it by 44.4%. Crude CFS showed activity even after three months of storage below -20°C and boiling for 10 min, whereas it loses 100% of its antimicrobial activity after enzymatic treatments of lipase, trypsin and papain. The production of antimicrobial compounds and broad spectrum of antimicrobial activity against all tested pathogens suggested that the strain DK1-SA11 can be used as a source for probiotics, synbiotics and antibiotics.

Designing and implementation of non-invasive methods for glucose monitoring in blood is main focus of biomedical scientists to provide a relief from skin puncturing of diabete patient. The objective of this research work is to investigate the shape deformations and the aggregation of red blood cells (RBCs) in the human blood after addition of three different analytes i) (0mM-400mM: Range) of glucose (C(6)H(12)O(6)), ii) (0mM-400mM: range) of pure salt (NaCl) and iii) (0mM- 350mM: range) of pure water (H(2)O). We have observed that the changes in the shape of individual cells from biconcave discs to spherical shapes and eventually the lysis of the cells at optimum concentration of glucose, salts and pure water. This demonstration also provides a base line to facilitate diabetes during partial diagnosis and monitoring of the glucose levels qualitatively both in research laboratories and clinical environment.

Acute coronary artery syndrome (ACS) is the major cause of mortality in Pakistan with genetic and environmental influence on the incidence of the disease. This case-control study was designed to find out if a correlation is existing between ACS and single nucleotide polymorphisms (SNPs) in DNA repair genes XPD [at codon 751, rs 13181 (Lys to Gln)] and XRCC1 [at codon 399, rs25487 (Arg to Gln); 280, rs25489 (Arg to His) and 194, rs 1799782 (Arg to Trp)] either individually or in various combination with each other (haplotype analysis). The objective of this study was to find out the association of various studied risk factors and serum lipid profile of the subjects with the disease, if any. PCR-RFLP method was used to determine genotype at specific codon in 221 subjects (115 ACS patients and 106 healthy controls) from Southern Punjab population. Genotypic and allelic frequency distribution among the cases and controls revealed that all the studied SNPs were not individually associated with the ACS. Haplotype analysis revealed that subjects having wild type combination of all three XRCC1 SNPs had greater susceptibility to ACS than any other studied genotypic combinations. Analysis of risk factors revealed that hypertension (P<0.001), age (P=0.05), education (P<0.001), gender (P<0.001), family history (P=0.005), smoking habit (P=0.002) and diabetes (P<0.001) were significantly associated with the incidence of ACS. Serum lipid profile analysis indicated that cholesterol level was significantly higher (P=0.048) in patients (161.5mg/dL) than controls (142.1mg/dL) while triglyceride remained unaffected (P=0.87) when compared between the two treatments.

Anti-microbial resistance burden and hazard associated with chemical treatment of infections demanded for new anti-microbial natural products. Marine associated microorganisms are the enormous source of bioactive compounds. In this study we have isolated 272 marine bacteria among them 136 (50%) were antagonistic to at least one of the four pathogenic strains Listeria monocytogenes, Vibrio cholerae, E. coli and S. aureus. Only two strains exhibited antibacterial activity against all four test strains, which were identified by 16S rDNA sequencing as Bacillus sp. DK1-SA11 and Vibrio sp. DK6-SH8. Marine isolate DK1-SA11 has potential to resist boiling temperature and pH 2-12. Furthermore cell free extract (CFE) inhibited all test organisms including superbug MRSA and pathogenic yeast Candida albicans. Marine isolate Bacillus sp. DK1-SA11 could be a potential combatant for the battle of drugs and bugs.

This study was carried out during 2005-2008 to identify existing plant species visited by workers of honeybees for nectar and pollen collection in Dera Ismail Khan (D.I.Khan) District, Pakistan. The honeybee species investigated in the area were, rock bee (Apis dorsata F.), little bee (A. florea F.) and European honeybee (A. mellifera L.). A detailed list of 86 plant species both wild and cultivated was prepared, out of which 12 species, Phulai (Acacia modesta Wall.), Sarsoon (Brassica campestris L.), Kaghzi nimboo (Citrus aurantifolia L.), Khatta (C. medica L.), Malta (C. sinensis (L.) Osbeck.), Shisham (Dalbergia sissoo Roxb.), Date (Phoenix dactylifera L.), Barseen (Trifolium alexandrianum L.), Shaftal (T. resupinatum L), Makai (Zea mays L.), Ber (Zizyphus mauritiana Lam.) and Jher beri (Z. numularia (Burm. f.) Wight & Arn.,) were found as major sources (table 1) for the production of surplus honey in a year at different localities in the District. Among the minor sources various plant species were included (table 2). These plants fill the flowering gaps between the major sources in various parts of the year and help in continuous supply of food to honey bees.

Hepatitis B virus (HBV) affects more than 350 million people worldwide and is a leading cause of morbidity and mortality in developing countries like Pakistan. Lamivudine has potential to inhibit hepatitis B virus (HBV) replication but long term lamivudine treatment results in mutations in YMDD region of HBV, making this therapy ineffective. In this study, we have optimized a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) based protocol to detect two mutations in HBV DNA polymerase gene (at codon 528 and 552) in chronic hepatitis patients, without any prior lamivudine treatment. HBV genome was extracted and tested by PCR-RFLP for detection of mutations in polymerase gene. Variations in HBV genome were not detected in enrolled patients confirming that lamivudine can be used to treat chronic Hepatitis B in these patients. Several studies have reported the natural occurrence of mutation in YMDD motif of polymerase gene in chronic hepatitis B patients, not treated with lamivudine, but these mutants were not detected in Pakistani lamivudine-untreated chronic hepatitis B patients.

Colorectal cancers (CRC) are highly prevalent cancer all over the world and need appropriate and timely prevention and treatment. Since years it has been argued that antioxidant vitamins have a potential role in the prevention of several neoplasm including colorectal cancer though the answer remained controversial. Most of the observational studies in past have shown that Vitamin E has some protective effect in the primary prevention of colorectal cancer, however its exact role is not yet established. On the other hand recently conducted experimental studies have shown variable results regarding the role of vitamin E in preventing colorectal cancers. Thus this review was conducted to study the role of vitamin E in preventing colorectal neoplasm. This review study was conducted from September 2008 to February 2009. We searched multiple electronic sources including (PUBMED) MEDLINE, Cochrane Database for identifying existing Systematic Reviews, OVID data base and other library sources to identify relevant studies for this review. Data was collected using data extraction form. Meta analysis was performed in Review Manager version 4.3. We identified four trials on vitamin E role for primary prevention of CRC, includes 94069 participants (47029 in vitamin E Vs 47040 in placebo), aged 40 years or above, who were randomized to vitamin E supplement versus placebo. The outcome measure in our review was incidence of colorectal cancer in the follow up period of 7 to 10 years. We found no sufficient evidence of vitamin E role for decreasing risk of CRC incidence (RR: 0.89, CI: 0.76, 1.05; p-value = 0.18). It has been identified in the review that Vitamin E does not have protective role in the prevention of colorectal cancer. Further studies on diverse population are required to determine the role vitamin E for the primary prevention of colorectal cancer.

Levels of various lipoprotein subclasses can provide useful information on coronary artery disease (CAD) risk status, even when traditional risk factors are known. Elevated serum level of lipids, lipoproteins and lipoprotein (a) have been identified as risk factor for atherosclerosis resulting in coronary artery disease, cerebrovascular disease, peripheral vascular disease and venous thromboembolism. The objective of this study is to evaluate the levels of serum lipids, lipoprotein and lipoprotein (a) in-patients suffering from documented coronary artery disease and normal healthy subjects. A case control study was conducted to evaluate the levels of lipid, lipoprotein and lipoprotein (a) in patients from our local population suffering from documented coronary artery disease. The present study was earned out in Dr. HMI Institute of Pharmacology & Herbal Sciences in collaboration with National Institute of Cardiovascular Diseases, Karachi from January 2001 to June 2001. One hundred and fifty persons were included in this case control study. Out of these one hundred were patients suffering from documented (coronary angiography done 4 month before the study) coronary artery disease (mean age 46.38 +/- 1.38, 18 females and 82 males). 50 normal healthy subjects (mean age, 44.86 +/- 1.15, 2 females and 48 males) had no history of IHD ever before. Their serum lipid, lipoprotein and lipoprotein (a) were estimated and compared.

Bergenia ciliata was subjected to antibacterial analysis. A battery of assays were performed on different extracts of Bergenia ciliata for antibacterial activities. Antibacterial effects of ethanol, hexane, ethyl acetate, chloroform, butanol and aqueous extracts of the roots and leaves extract of Bergenia ciliata exhibited various degree of inhibition activity. It was observed that root and leaves extract were promising against gram positive and gram negative bacteria viz. Bacillus subtilis, Bacillus megaterium and Pseudomonas aeruginosa. This study showed that Bergenia ciliata could inhibit selectively bacteria.

Bergenia ciliata was subjected to bioactivity analysis. The records of these investigations are described in this communication. A study made on the bioactivity analysis of medicinal herb Bergenia ciliata, which in folkloric medicine is used to cure hypoglycemic activity. A battery of assays was performed on different extracts of Bergenia ciliata which include hypoglycemic activities, toxic evaluations such as acute systemic and intracutaneous toxicity as well hemolysis test. Bergenia ciliata has been employed in folklore medicine to treat symptoms of diabetes mellitus. All the extracts except chloroform extract of root and leaves of Bergenia ciliata were found to possess hypoglycemic activity in Streptozotocin (STZ) treated rats. Therefore the plant can be classified as hypoglycemic, hypoglycemic activity in experimental diabetes ranging from 40-70% of its onset to reduce blood glucose level. The toxicological investigations of Bergenia ciliata with particular reference to acute systematic toxicity and intracutaneous toxicity in experimental animals displayed that it elicit severe toxicity. The symptoms of toxicity in intracutaneous test showed erythema and edema whereas assessment of acute systemic toxicity frequently observed breathing problem and initiations of diarrhea with blood in stool of experimental model and caused gastero-intestinal syndrome. Bergenia ciliata can produce toxicity suggesting a role in certain diseases. It is therefore, premature to speculate about mechanism of effect until toxin(s) is unequivocally identified. The hemolysis test on the extract of Bergenia ciliata was almost devoid of activity.